ABSTRACT
Quorum sensing (QS) inhibition stands out as an innovative therapeutic strategy for combating infections caused by drug-resistant pathogens. In this study, we assessed the potential of 3-(2-isocyanobenzyl)-1H-indole derivatives as novel quorum sensing inhibitors (QSIs). Initial screenings of their QS inhibitory activities were conducted against Pseudomonas aeruginosa PAO1 and Chromobacterium violaceum CV026. Notably, six 3-(2-isocyanobenzyl)-1H-indole derivatives (4, 12, 25, 28, 32, and 33) exhibited promising QS, biofilms, and pyocyanin inhibitory activities under minimum inhibitory concentrations (MICs) against P. aeruginosa PAO1. Among them, 3-(2-isocyano-6-methylbenzyl)-1H-indole (IMBI, 32) emerged as the most promising candidate, demonstrating superior biofilm and pyocyanin inhibition. Further comprehensive studies revealed that derivative 32 at 25 µg mL-1 inhibited biofilm formation by 70% against P. aeruginosa PAO1, as confirmed by scanning electron microscopy (SEM). Additionally, derivative 32 substantially increased the susceptibility of mature biofilms, leading to a 57% destruction of biofilm architecture. In terms of interfering with virulence factors in P. aeruginosa PAO1, derivative 32 (25 µg mL-1) displayed remarkable inhibitory effects on pyocyanin, protease, and extracellular polysaccharides (EPS) by 73%, 51%, and 37%, respectively, exceeding the positive control resveratrol (RSV). Derivative 32 at 25 µg mL-1 also exhibited effective inhibition of swimming and swarming motilities. Moreover, it downregulated the expressions of QS-related genes, including lasI, lasR, rhlI, rhlR, pqsR, sdhB, sucD, sodB, and PA5439, by 1.82- to 10.87-fold. Molecular docking, molecular dynamics simulations (MD), and energy calculations further supported the stable binding of 32 to LasR, RhlI, RhlR, EsaL, and PqsR antagonizing the expression of QS-linked traits. Evaluation of the toxicity of derivative 32 on HEK293T cells via CCK-8 assay demonstrated low cytotoxicity. Overall, this study underscores the efficacy of derivative 32 in inhibiting virulence factors in P. aeruginosa. Derivative 32 emerges as a potential QSI for controlling P. aeruginosa PAO1 infections in vitro and an anti-biofilm agent for restoring or enhancing drug sensitivity in drug-resistant pathogens.
ABSTRACT
Quantum-dot light-emitting diodes (QLEDs) are solution-processed electroluminescence devices with great potential as energy-saving, large-area, and low-cost display and lighting technologies. Ideally, the organic hole-transport layers (HTLs) in QLEDs should simultaneously deliver efficient hole injection and transport, effective electron blocking, and robust electrochemical stability. However, it is still challenging for a single HTL to fulfill all these stringent criteria. Here, we demonstrate a general design of doping-bilayer polymer-HTL architecture for stabilizing high-efficiency QLEDs. We show that the bilayer HTLs combining the electrochemical-stable polymer and the electron-blocking polymer unexpectedly increase the hole injection barrier. We mitigated the problem by p-doping of the underlying sublayer of the bilayer HTLs. Consequently, green QLEDs with an unprecedented maximum luminance of 1,340,000 cd m-2 and a record-long operational lifetime (T95 lifetime at an initial luminance of 1000 cd m-2 is 17,700 hours) were achieved. The universality of the strategy is examined in various polymer-HTL systems, providing a general route toward high-performance solution-processed QLEDs.
ABSTRACT
Anti/deicing coatings that combine active and passive methods can utilize various energy sources to achieve anti/deicing effects. However, poor photothermal or electrothermal performance and inevitable heat loss often reduce their anti/deicing efficiency. Herein, copper sulfide loaded activated biochar (AC@CuS) as photo/electric material, polydimethylsiloxane as hydrophobic component, thermally expandable microspheres as foaming agent, and an anti/deicing coating integrating thermal insulation, superhydrophobicity, photo/electrothermal effects was successfully constructed. Benefiting from the synergistic effect of superhydrophobicity and thermal insulation, the freezing time of water droplets on the coating surface is extended from 150 to 2140 s, showing excellent passive anti-icing performance. AC@CuS exhibits photo/electrothermal effects, and porous expanded microspheres reduce heat loss, which endows the coating with desirable photo/electrothermal conversion performance. Under the conditions of 0.2 W/cm2 electric power density (EPD) and 0.1 W/cm2 optical power density (OPD), the temperature of the coating increases from 24 to 96.4 and 113 °C, respectively. Interestingly, with a coheating of 0.05 W/cm2 weaker OPD and 0.05 W/cm2 lower EPD, the ice on the coating surface can be quickly melted in 2.5 min, showing synergistic deicing performance. In addition, the WCA of the prepared coating remains above 150° after mechanical damage, rain impact, UV irradiation, chemical corrosion, and high-temperature treatment, and good superhydrophobic durability ensures the anti/deicing durability of the coating.
ABSTRACT
Photothermal modulation of neural activity offers a promising approach for understanding brain circuits and developing therapies for neurological disorders. However, the low neuron selectivity and inefficient light-to-heat conversion of existing photothermal nanomaterials significantly limit their potential for neuromodulation. Here, we report that graphdiyne (GDY) can be developed into an efficient neuron-targeted photothermal transducer for in vivo modulation of neuronal activity through rational surface functionalization. We functionalize GDY with polyethylene glycol (PEG) through noncovalent hydrophobic interactions, followed by antibody conjugation to specifically target the temperature-sensitive transient receptor potential cation channel subfamily V member 1 (TRPV1) on the surface of neural cells. The nanotransducer not only exhibits high photothermal conversion efficiency in the near-infrared region but also shows great TRPV1-targeting capability. This enables photothermal activation of TRPV1, leading to neurotransmitter release in cells and modulation of neural firing in living mice. With its precision and selectivity, the GDY-based transducer provides an innovative avenue for understanding brain function and developing therapeutic strategies for neurodegenerative diseases.
Subject(s)
Neurons , TRPV Cation Channels , Animals , TRPV Cation Channels/metabolism , Neurons/metabolism , Mice , Humans , Graphite/chemistry , Graphite/pharmacology , Polyethylene Glycols/chemistry , TransducersABSTRACT
The SPRY domain-containing SOCS box proteins SPSB1, SPSB2, and SPSB4 utilize their SPRY/B30.2 domain to interact with a short region in the N-terminus of inducible nitric oxide synthase (iNOS), and recruit an E3 ubiquitin ligase complex to polyubiquitinate iNOS, resulting in the proteasomal degradation of iNOS. Inhibitors that can disrupt the endogenous SPSB-iNOS interactions could be used to augment cellular NO production, and may have antimicrobial and anticancer activities. We previously reported the rational design of a cyclic peptide inhibitor, cR8, cyclo(RGDINNNV), which bound to SPSB2 with moderate affinity. We, therefore, sought to develop SPSB inhibitors with higher affinity. Here, we show that cyclic peptides cR7, cyclo(RGDINNN), and cR9, cyclo(RGDINNNVE), have ~6.5-fold and ~2-fold, respectively, higher SPSB2-bindng affinities than cR8. We determined high-resolution crystal structures of the SPSB2-cR7 and SPSB2-cR9 complexes, which enabled a good understanding of the structure-activity relationships for these cyclic peptide inhibitors. Moreover, we show that these cyclic peptides displace full-length iNOS from SPSB2, SPSB1, and SPSB4, and that their inhibitory potencies correlate well with their SPSB2-binding affinities. The strongest inhibition was observed for cR7 against all three iNOS-binding SPSB proteins.
Subject(s)
Peptides, Cyclic , Peptides, Cyclic/chemistry , Peptides, Cyclic/pharmacology , Humans , Suppressor of Cytokine Signaling Proteins/chemistry , Suppressor of Cytokine Signaling Proteins/metabolism , Nitric Oxide Synthase Type II/antagonists & inhibitors , Nitric Oxide Synthase Type II/metabolism , Nitric Oxide Synthase Type II/chemistry , Oligopeptides/chemistry , Oligopeptides/pharmacology , Protein Binding , Structure-Activity RelationshipABSTRACT
The surge in lithium-ion batteries has heightened concerns regarding metal resource depletion and the environmental impact of spent batteries. Battery recycling has become paramount globally, but conventional techniques, while effective at extracting transition metals like cobalt and nickel from cathodes, often overlook widely used spent LiFePO4 due to its abundant and low-cost iron content. Direct regeneration, a promising approach for restoring deteriorated cathodes, is hindered by practicality and cost issues despite successful methods like solid-state sintering. Hence, a smart prelithiation separator based on surface-engineered sacrificial lithium agents is proposed. Benefiting from the synergistic anionic and cationic redox, the prelithiation separator can intelligently release or intake active lithium via voltage regulation. The staged lithium replenishment strategy was implemented, successfully restoring spent LiFePO4's capacity to 163.7â mAh g-1 and a doubled life. Simultaneously, the separator can absorb excess active lithium up to approximately 600â mAh g-1 below 2.5â V to prevent over-lithiation of the cathode This innovative, straightforward, and cost-effective strategy paves the way for the direct regeneration of spent batteries, expanding the possibilities in the realm of lithium-ion battery recycling.
ABSTRACT
Doping plays a crucial role in modulating and enhancing the performance of organic semiconductor (OSC) devices. In this study, the critical role of dopants is underscored in shaping the morphology and structure of OSC films, which in turn profoundly influences their properties. Two dopants, trityl tetrakis(pentafluorophenyl) (TrTPFB) and N,N-dimethylanilinium tetrakis(pentafluorophenyl)borate (DMA-TPFB), are examined for their doping effects on poly(3-hexylthiophene) (P3HT) and PBBT-2T host OSCs. It is found that although TrTPFB exhibits higher doping efficiency, OSCs doped with DMA-TPFB achieve comparable or even enhanced electrical conductivity. Indeed, the electrical conductivity of DMA-TPFB-doped P3HT reaches over 67 S cm-1, which is a record-high value for mixed-solution-doped P3HT. This can be attributed to DMA-TPFB inducing a higher degree of crystallinity and reduced structural disorder. Moreover, the beneficial impact of DMA-TPFB on the OSC films' morphology and structure results in superior thermoelectric performance in the doped OSCs. These findings highlight the significance of dopant-induced morphological and structural considerations in enhancing the film characteristics of OSCs, opening up a new avenue for optimization of dopant performance.
ABSTRACT
Reliably discerning real human faces from fake ones, known as antispoofing, is crucial for facial recognition systems. While neuromorphic systems offer integrated sensing-memory-processing functions, they still struggle with efficient antispoofing techniques. Here we introduce a neuromorphic facial recognition system incorporating multidimensional deep ultraviolet (DUV) optoelectronic synapses to address these challenges. To overcome the complexity and high cost of producing DUV synapses using traditional wide-bandgap semiconductors, we developed a low-temperature (≤70 °C) solution process for fabricating DUV synapses based on PEA2PbBr4/C8-BTBT heterojunction field-effect transistors. This method enables the large-scale (4-in.), uniform, and transparent production of DUV synapses. These devices respond to both DUV and visible light, showing multidimensional features. Leveraging the unique ability of the multidimensional DUV synapse (MDUVS) to discriminate real human skin from artificial materials, we have achieved robust neuromorphic facial recognition with antispoofing capability, successfully identifying genuine human faces with an accuracy exceeding 92%.
ABSTRACT
Selective and nondisruptive in vivo neurochemical monitoring within the central nervous system has long been a challenging endeavor. We introduce a new sensing approach that integrates neurocompatible galvanic redox potentiometry (GRP) with customizable phosphorothioate aptamers to specifically probe dopamine (DA) dynamics in live rat brains. The aptamer-functionalized GRP (aptGRP) sensor demonstrates nanomolar sensitivity and over a 10-fold selectivity for DA, even amidst physiological levels of major interfering species. Notably, conventional sensors without the aptamer modification exhibit negligible reactivity to DA concentrations exceeding 20 µM. Critically, the aptGRP sensor operates without altering neuronal activity, thereby permitting real-time, concurrent recordings of both DA flux and electrical signaling in vivo. This breakthrough establishes aptGRP as a viable and promising framework for the development of high-fidelity sensors, offering novel insights into neurotransmission dynamics in a live setting.
Subject(s)
Aptamers, Nucleotide , Brain , Dopamine , Potentiometry , Animals , Aptamers, Nucleotide/chemistry , Dopamine/analysis , Rats , Potentiometry/methods , Potentiometry/instrumentation , Brain/metabolism , Biosensing Techniques/methods , Rats, Sprague-Dawley , MaleABSTRACT
The formation of a solid electrolyte interphase on carbon anodes causes irreversible loss of Na+ ions, significantly compromising the energy density of Na-ion full cells. Sodium compensation additives can effectively address the irreversible sodium loss but suffer from high decomposition voltage induced by low electrochemical activity. Herein, we propose a universal electrocatalytic sodium compensation strategy by introducing a carbon nanotube (CNT)/MnO2 catalyst to realize full utilization of sodium compensation additives at a much-reduced decomposition voltage. The well-organized CNT/MnO2 composite with high catalytic activity, good electronic conductivity, and abundant reaction sites enables sodium compensation additives to decompose at significantly reduced voltages (from 4.40 to 3.90 V vs Na+/Na for sodium oxalate, 3.88 V for sodium carbonate, and even 3.80 V for sodium citrate). As a result, sodium oxalate as the optimal additive achieves a specific capacity of 394 mAh g-1, almost reaching its theoretical capacity in the first charge, increasing the energy density of the Na-ion full cell from 111 to 158 Wh kg-1 with improved cycle stability and rate capability. This work offers a valuable approach to enhance sodium compensation efficiency, promising high-performance energy storage devices in the future.
ABSTRACT
Intercellular communication often relies on exosomes as messengers and is critical for cancer metastasis in hypoxic tumor microenvironment. Some circular RNAs (circRNAs) are enriched in cancer cell-derived exosomes, but little is known about their ability to regulate intercellular communication and cancer metastasis. Here, by systematically analyzing exosomes secreted by non-small cell lung cancer (NSCLC) cells, a hypoxia-induced exosomal circPLEKHM1 is identified that drives NSCLC metastasis through polarizing macrophages toward to M2 type. Mechanistically, exosomal circPLEKHM1 promoted PABPC1-eIF4G interaction to facilitate the translation of the oncostatin M receptor (OSMR), thereby promoting macrophage polarization for cancer metastasis. Importantly, circPLEKHM1-targeted therapy significantly reduces NSCLC metastasis in vivo. circPLEKHM1 serves as a prognostic biomarker for metastasis and poor survival in NSCLC patients. This study unveils a new circRNA-mediated mechanism underlying how cancer cells crosstalk with macrophages within the hypoxic tumor microenvironment to promote metastasis, highlighting the importance of exosomal circPLEKHM1 as a prognostic biomarker and therapeutic target for lung cancer metastasis.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Exosomes , Lung Neoplasms , Macrophages , RNA, Circular , Tumor Microenvironment , Animals , Humans , Mice , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/metabolism , Carcinoma, Non-Small-Cell Lung/pathology , Cell Line, Tumor , Disease Models, Animal , Exosomes/metabolism , Exosomes/genetics , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Macrophages/metabolism , Neoplasm Metastasis/genetics , RNA, Circular/genetics , RNA, Circular/metabolism , Tumor Microenvironment/genetics , Mice, NudeABSTRACT
Vitamin C (Vc) plays a pivotal role in a series of pathological processes, such as tumors, immune diseases, and neurological disorders. However, its therapeutic potential for tinnitus management remains unclear. In this study, we find that Vc relieves tinnitus in noise-exposed rats. In the 7-day therapy groups, spontaneous firing rate (SFR) increases from 1.17 ± 0.10 Hz to 1.77 ± 0.15 Hz after noise exposure. Vc effectively reduces the elevated SFR to 0.99 ± 0.07 and 0.55 ± 0.05 Hz at different doses. The glutamate level in auditory cortex of noise-exposed rats (3.78 ± 0.42 µM) increases relative to that in the control group (1.34 ± 0.22 µM). High doses of Vc (500 mg/kg/day) effectively reduce the elevated glutamate levels (1.49 ± 0.28 µM). Mechanistic studies show that the expression of glutamate transporter 1 (GLT-1) is impaired following noise exposure and that Vc treatment effectively restores GLT-1 expression in the auditory cortex. Meanwhile, the GLT-1 inhibitor, dl-threo-beta-benzyloxyaspartic acid (dl-TBOA), invalidates the protection role of Vc. Our finding shows that Vc substantially enhances glutamate clearance by upregulating GLT-1 and consequently alleviates noise-induced tinnitus. This study provides valuable insight into a novel biological target for the development of therapeutic interventions that may prevent the onset of tinnitus.
Subject(s)
Auditory Cortex , Tinnitus , Rats , Animals , Auditory Cortex/metabolism , Ascorbic Acid/pharmacology , Ascorbic Acid/metabolism , Neuroprotection , Tinnitus/drug therapy , Tinnitus/metabolism , Glutamic Acid/metabolism , Disease Models, Animal , Amino Acid Transport System X-AG/metabolism , Excitatory Amino Acid Transporter 2/metabolismABSTRACT
To assess the strain resources and address production challenges in Ganoderma cultivation. 150 Ganoderma strains were collected from 13 provinces in China. A comparative analysis of agronomic traits and effective components was conducted. Among the 150 strains, key agronomic traits measured were: average stipe diameter (15.92 mm), average stipe length (37.46 mm), average cap horizontal diameter (94.97 mm), average cap vertical diameter (64.21 mm), average cap thickness (15.22 mm), and average fruiting body weight (14.30 g). Based on these agronomic traits, four promising strains, namely, L08, L12, Z21, and Z39, were recommended for further cultivation and breeding. The average crude polysaccharide content ranged from 0.048% to 0.977%, and triterpenoids ranged from 0.804% to 2.010%. In addition, 73 triterpenoid compounds were identified, constituting 47.1% of the total compounds. Using a distance discrimination method, the types, and relative contents of triterpenoid compounds in 150 Ganoderma strains were classified, achieving 98% accuracy in G. lingzhi identification. The 16 triterpenoid components used for G. lingzhi identification included oleanolic acid, ursolic acid, 3ß-acetoxyergosta-7,22-dien-5α-ol, ganoderic acid DM, ganoderiol B, ganorderol A, ganoderic acid GS-1, tsugaric acid A, ganoderic acid GS-2, ganoderenic acid D, ganoderic acid Mf, ganoderic acid A, ganoderic acid K, ganoderic acid V, ganoderic acid G, and leucocontextin J. This study provides valuable insights for exploring and utilizing Ganoderma resources and for the development of new varieties.
Subject(s)
Agaricales , Agaricus , Antineoplastic Agents , Ganoderma , Reishi , Triterpenes , Triterpenes/analysis , ChinaABSTRACT
O3-type layered oxide cathodes (NaxTMO2) for sodium-ion batteries (SIBs) have attracted significant attention as one of the most promising potential candidates for practical energy storage applications. The poor Na+ diffusion kinetics is, however, one of the major obstacles to advancing large-scale practical application. Herein, we report bismuth-doped O3-NaNi0.5Mn0.5O2 (NMB) microspheres consisting of unique primary nanoplatelets with the radially oriented {010} active lattice facets. The NMB combines the advantages of the oriented and exposed electrochemical active planes for direct paths of Na+ diffusion, and the thick primary nanoplatelets for less surface parasitic reactions with the electrolyte. Consequently, the NMB cathode exhibits a long-term stability with an excellent capacity retention of 72.5% at 1C after 300 cycles and an enhanced rate capability at a 0.1C to 10C rate (1C = 240 mA g-1). Furthermore, the enhancement is elucidated by the small volume change, thin cathode-electrolyte-interphase (CEI) layer, and rapid Na+ diffusion kinetics. In particular, the radial orientation-based Bi-doping strategy is demonstrated to be effective at boosting electrochemical performance in other layered oxides (such as Bi-doped NaNi0.45Mn0.45Ti0.1O2 and NaNi1/3Fe1/3Mn1/3O2). The results provide a promising strategy of utilizing the advantages of the oriented active facets of primary platelets and secondary particles to develop high-rate layered oxide cathodes for SIBs.
ABSTRACT
The intermitochondrial cement (IMC) and chromatoid body (CB) are posited as central sites for piRNA activity in mice, with MIWI initially assembling in the IMC for piRNA processing before translocating to the CB for functional deployment. The regulatory mechanism underpinning MIWI translocation, however, has remained elusive. We unveil that piRNA loading is the trigger for MIWI translocation from the IMC to CB. Mechanistically, piRNA loading facilitates MIWI release from the IMC by weakening its ties with the mitochondria-anchored TDRKH. This, in turn, enables arginine methylation of MIWI, augmenting its binding affinity for TDRD6 and ensuring its integration within the CB. Notably, loss of piRNA-loading ability causes MIWI entrapment in the IMC and its destabilization in male germ cells, leading to defective spermatogenesis and male infertility in mice. Collectively, our findings establish the critical role of piRNA loading in MIWI translocation during spermatogenesis, offering new insights into piRNA biology in mammals.
Subject(s)
Argonaute Proteins , Germ Cell Ribonucleoprotein Granules , Piwi-Interacting RNA , Animals , Male , Mice , Argonaute Proteins/metabolism , Germ Cells/metabolism , Mammals/genetics , Mitochondria/metabolism , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Spermatogenesis/genetics , Testis/metabolismABSTRACT
Redox potentiometry has emerged as a new platform for in vivo sensing, with improved neuronal compatibility and strong tolerance against sensitivity variation caused by protein fouling. Although enzymes show great possibilities in the fabrication of selective redox potentiometry, the fabrication of an enzyme electrode to output open-circuit voltage (EOC) with fast response remains challenging. Herein, we report a concept of novel enzymatic galvanic redox potentiometry (GRP) with improved time response coupling the merits of the high selectivity of enzyme electrodes with the excellent biocompatibility and reliability of GRP sensors. With a glucose biosensor as an illustration, we use flavin adenine dinucleotide-dependent glucose dehydrogenase as the recognition element and carbon black as the potential relay station to improve the response time. We find that the enzymatic GRP biosensor rapidly responds to glucose with a good linear relationship between EOC and the logarithm of glucose concentration within a range from 100 µM to 2.65 mM. The GRP biosensor shows high selectivity over O2 and coexisting neurochemicals, good reversibility, and sensitivity and can in vivo monitor glucose dynamics in rat brain. We believe that this study will pave a new platform for the in vivo potentiometric biosensing of chemical events with high reliability.
Subject(s)
Biosensing Techniques , Glucose Oxidase , Potentiometry , Reproducibility of Results , Glucose Oxidase/metabolism , Electrodes , Glucose , Oxidation-Reduction , Glucose 1-Dehydrogenase/metabolismABSTRACT
This study aimed to investigate the species diversity and genetic differentiation of the genome of the main cultivated strains of Ganoderma in China. Population genomics analysis was conducted based on 150 cultivated strains of Ganoderma collected nationwide. The results indicated that the main species currently cultivated in China were Ganoderma sichuanense and Ganoderma lucidum, with a minor proportion of Ganoderma sessile, Ganoderma weberianum, Ganoderma sinense, Ganoderma gibbosum and Ganoderma australe. A total of 336,506 high-quality single nucleotide polymorphism (SNP) loci were obtained through population evolution analysis. The Fst values were calculated using a 5-kb sliding window, which ranged from 0.11 to 0.74. This suggests varying degrees of genetic differentiation between populations and genetic exchange among varieties. On this basis, the genes related to the stipe length, cap color and branch phenotypes of Ganoderma were excavated, and the region with the top 1% ZFst value region was used as a candidate region. A total of 137, 270 and 222 candidate genes were identified in the aforementioned 3 phenotypes, respectively. Gene annotation revealed that genes associated with stipe length were mainly related to cell division and differentiation, including proteins such as Nse4 protein and DIM1 protein. The genes related to Ganoderma red color were mainly related to the metabolism of tryptophan and flavonoids. The genes related to the branch were mainly related to cytokinin synthesis, ABC transporter and cytochrome P450. This study provided 150 valuable genome resequencing data in assessing the diversity and genetic differentiation of Ganoderma and laid a foundation for agronomic trait analysis and the development of new varieties of Ganoderma.
Subject(s)
Ganoderma , Genetics, Population , Genetic Drift , Ganoderma/genetics , ChinaABSTRACT
OBJECTIVE: Ferroptosis is a reactive oxygen species (ROS)- and iron-dependent form of non-apoptotic cell death process. Previous studies have demonstrated that ferroptosis participates in the development of inflammatory arthritis. However, the role of ferroptosis in rheumatoid arthritis (RA) inflammatory hypoxic joints remains unclear. This study sought to explore the underlying mechanism of ferroptosis on lipopolysaccharide (LPS)-induced RA fibroblast-like synoviocytes (FLSs). METHODS: FLSs, isolated from patients with RA, were treated with LPS and ferroptosis inducer (erastin and RSL-3), and ferroptosis inhibitor (Fer-1 and DFO), respectively. The cell viability was measured by CCK-8. The cell death was detected by flow cytometer. The proteins level were tested by Western blot. The cytosolic ROS and lipid peroxidation were determined using DCFH-DA and C11-BODIPY581/591 fluorescence probes, respectively. The small interfering RNA (siRNA) was used to knock down related proteins. The levels of malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), iron, inflammatory cytokines (IL6 and IL8), and LDH were analyzed by commercial kits. RESULTS: Ferroptosis was activated by LPS in RA FLS with increased cellular damage, ROS and lipid peroxidation, intracellular Fe and IL8, which can be further amplified by ferroptosis inducer (erastin and RSL-3) and inhibited by ferroptosis inhibitor (Fer-1 and DFO). Mechanistically, LPS triggered ferroptosis via NCOA4-mediated ferritinophagy in RA FLSs, and knockdown of NCOA4 strikingly prevent the process of ferroptosis. Intriguingly, LPS-induced RA FLSs became insensitive to ferroptosis and NCOA4-mediated ferritinophagy under hypoxia compared with normoxia. Knockdown of HIF-1α reverted ferroptosis and ferritinophagy evoking by LPS-induced RA FLSs inflammation under hypoxia. In addition, low dose of auranofin (AUR) induced re-sensitization of ferroptosis and ferritinophagy through inhibiting the expression of HIF-1α under hypoxia. CONCLUSIONS: NCOA4-mediated ferritinophagy was a key driver of ferroptosis in inflammatory RA FLSs. The suppression of NCOA4-mediated ferritinophagy protected RA FLSs from ferroptosis in LPS-induced inflammation under hypoxia. Targeting HIF-1α/NCOA4 and ferroptosis could be an effective and valuable therapeutic strategy for synovium hyperplasia in the patients with RA.
Subject(s)
Arthritis, Rheumatoid , Ferroptosis , Synoviocytes , Humans , Lipopolysaccharides/pharmacology , Reactive Oxygen Species/metabolism , Interleukin-8/metabolism , Arthritis, Rheumatoid/metabolism , Inflammation/chemically induced , Inflammation/metabolism , Hypoxia/metabolism , Transcription Factors/metabolism , RNA, Small Interfering/genetics , Fibroblasts/metabolism , Iron/metabolism , Nuclear Receptor Coactivators/genetics , Nuclear Receptor Coactivators/metabolismABSTRACT
BACKGROUND: Clinical severity scores, such as acute physiology, age, chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA), Pitt Bacteremia Score (PBS), and European Confederation of Medical Mycology Quality (EQUAL) score, may not reliably predict candidemia prognosis owing to their prespecified scorings that can limit their adaptability and applicability. OBJECTIVES: Unlike those fixed and prespecified scorings, we aim to develop and validate a machine learning (ML) approach that is able to learn predictive models adaptively from available patient data to increase adaptability and applicability. METHODS: Different ML algorithms follow different design philosophies and consequently, they carry different learning biases. We have designed an ensemble meta-learner based on stacked generalisation to integrate multiple learners as a team to work at its best in a synergy to improve predictive performances. RESULTS: In the multicenter retrospective study, we analysed 512 patients with candidemia from January 2014 to July 2019 and compared a stacked generalisation model (SGM) with APACHE II, SOFA, PBS and EQUAL score to predict the 14-day mortality. The cross-validation results showed that the SGM significantly outperformed APACHE II, SOFA, PBS, and EQUAL score across several metrics, including F1-score (0.68, p < .005), Matthews correlation coefficient (0.54, p < .05 vs. SOFA, p < .005 vs. the others) and the area under the curve (AUC; 0.87, p < .005). In addition, in an independent external test, the model effectively predicted patients' mortality in the external validation cohort, with an AUC of 0.77. CONCLUSIONS: ML models show potential for improving mortality prediction amongst patients with candidemia compared to clinical severity scores.
Subject(s)
Bacteremia , Candidemia , Humans , Organ Dysfunction Scores , APACHE , Retrospective Studies , Candidemia/diagnosis , Feasibility Studies , Prognosis , Machine Learning , ROC Curve , Intensive Care UnitsABSTRACT
OBJECTIVE: This study aimed to assess the impact of a hydroethanolic extract of walnut green husks (WGH) on rumen fermentation and the diversity of bacteria, methanogenic archaea, and fungi in sheep fed a high-concentrate diet. METHODS: Five healthy small-tailed Han ewes with permanent rumen fistula were selected and housed in individual pens. This study adopted a self-controlled and crossover design with a control period and an experimental period. During the control period, the animals were fed a basal diet (with a ratio of concentrate to roughage of 65:35), while during the treatment period, the animals were fed the basal diet supplemented with 0.5% hydroethanolic extract of WGH. Fermentation parameters, digestive enzyme activities, and microbial diversity in rumen fluid were analyzed. RESULTS: Supplementation of hydroethanolic extract of WGH had no significant effect on feed intake, concentrations of total volatile fatty acids, isovalerate, ammonia nitrogen, and microbial protein (p>0.05). However, the ruminal pH, concentrations of acetate, butyrate and isobutyrate, the ratio of acetate to propionate, protozoa count, and the activities of filter paper cellulase and cellobiase were significantly increased (p<0.05), while concentrations of propionate and valerate were significantly decreased (p<0.05). Moreover, 16S rRNA gene sequencing revealed that the relative abundance of rumen bacteria Christensenellaceae R7 group, Saccharofermentans, and Ruminococcaceae NK4A214 group were significantly increased, while Ruminococcus gauvreauii group, Prevotella 7 were significantly decreased (p<0.05). The relative abundance of the fungus Pseudomonas significantly increased, while Basidiomycota, Fusarium, and Alternaria significantly decreased (p<0.05). However, there was no significant change in the community structure of methanogenic archaea. CONCLUSION: Supplementation of hydroethanolic extract of WGH to a high-concentrate diet improved the ruminal fermentation, altered the structure of ruminal bacterial and fungal communities, and exhibited beneficial effects in alleviating subacute rumen acidosis of sheep.